|Public on Mar 07, 2014
|99WT SERTOLI CELLS
|Wildtype sertoli cells
mouse age: 5 months
proband sex: Male
tissue: cell isolation from testis
|mice were euthanized with carbon dioxide and the whole testis was removed. Germ cells and Sertoli cells were isolated.
|animals were housed at the mouse facility of the ENI, University Göttingen
|mRNA was isolated from the germ and Sertoli cell population, which were isolated from murine WT and TAp73KO testis. Cell samples were transferred into trizol buffer, homogenized and mRNA was isolated in a single-step acid guanidinium thiocyanate-phenol-chloroform extraction following manufacturer´s instructions (http://tools.invitrogen.com/content/sfs/manuals/trizol_reagent.pdf). RNA-precipitation was done with Ethanol. RNA was diluted in nuclease-free water and concentrations were determined using a Nanodrop spectrophotometer.
|For each array 200 ng RNA was processed and Cy3 labeled by the Low Input Quick Amp Labeling Kit (Agilent Technologies).
|Hybridization was performed in the hybridization oven (Agilent Technologies, 17h, 10 rpm, 65˚C)
|Cy3 intensities were detected by a one-color microarray scanner (G2505C, Agilent Technologies) at 5 µm resolution
|Data were quantile normalized and log2 transformed using R and Bioconductor package limma.
|Nov 06, 2013
|Last update date
|Mar 07, 2014
|Department of Human Genetics
|NGS Integrative Genomics
|TAp73 is essential for germ cell adhesion and maturation in testis